GB1577540A - Derivatives of 4,6-di-o-(aminoglycosyl)-1,3-dimaniocyclitols - Google Patents

Description

PATENT SPECIFICATION ( 11) 1 577 540

( 21) Application No 44599/77 ( 22) Filed 26 Oct 1977 14 ( 31) Convention Application No 736640 ( 1 ( 32) Filed 28 Oct 1976 in ( 33) United States of America (US) ( 44) Complete Specification published 22 Oct 1980 ( 51) INT CL 3 C 07 H 15/22 A 61 K 31/35 (C 07 H 15/22 C 07 D 309/08 309/16 309/20 309/28) ( 52) Index at acceptance C 2 C 1377 1671 1672 214 215 220 221 225 226 22 Y 246 247 253 255 25 Y 281 282 28 X 30 Y 313 314 31 Y 321 322 324 32 Y 337 339 340 342 34 Y 351 352 358 360 361 362 363 364 366 368 36 Y 37 X 390 395 39 Y 440 450 455 45 Y 47 X 509 50 Y 560 577 601 60 X 610 620 623 627 62 X 633 634 638 640 643 647 648 650 652 658 65 X 660 662 670 672 67 X 682 699 702 70 Y 71 Y 72 Y 774 777 778 801 80 Y AA BJ BL KA KB KH KQ LH LY MA NM NX QG RA WL ( 54) DERIVATIVES OF 4,6-DI-O-(AMINOGLYCOSYL)1,3-DIAMINOCYCLITOLS ( 71) We, SCHERICO LTD, Topferstrasse 5, Lucerne, Switzerland, a body corporate constituted under the laws of Switzerland, do hereby declare the invention, for which we pray that a patent may be granted to us, and the method by which it is to be performed, to be particularly described in and by the following

statement: 5

This invention relates to novel 2 '-unsubstituted derivatives of 4,6 di O (aminoglycosyl) 1,3 diaminocyclitols useful as antibacterial agents Further, this invention relates to pharmaceutical compositions comprising said 2 'unsubstituted derivatives and to methods for their manufacture.

Particularly, this invention relates to novel 2 '-unsubstituted derivatives of 4,6 10 di O (aminoglycosyl) 1,3 diaminocyclitol antibiotics including certain gentamicins, sisomicin, verdamicin, kanamycin A, Antibiotic JI-20 B, 6640 B, 66D, G-52, Mu-l, Mu-4, and further when the 1,3-diaminocyclitol has a 5hydroxyl group in the equatorial position, the 5-epi,5-deoxy, 5-epi-azido-5-deoxy, 5-epiamino-5-deoxy derivatives thereof and the l-N-derivatives of the foregoing 15 Known in the art are 4,6 di O (aminoglycosyl) 1,3 diaminocyclitols useful as antibacterial agents, all of which contain a hydroxyl or amino function or derivative thereof at the 2 '-position Antibacterial agents of this group include the gentamicins, sisomicin, verdamicin, the kanamycins and Antibiotics JI-20 B, 6640 B, 66-40 D, G-52, Mu-l and Mu-4 Also known in the art are the 1-Nalkyl and 1 20 N-acyl derivatives of the foregoing and the 5-epi derivatives of the foregoing, as well as the 5-epi-azido-5-deoxy and 5-epi-amino-5-deoxy derivatives all of which are also antibacterial agents.

By this invention novel 2 '-unsubstituted derivatives of the foregoing 4, 6 di O (aminoglycosyl) 1,3 diaminocyclitols are provided which are useful 25 antibacterial agents and which advantageously are cidal against certain microorganisms being resistant to the corresponding 2 ' substituted 4,6 di O (aminoglycosyl) 1,3 diaminocyclitols.

The compounds of this invention are 2 ' unsubstituted 4,6 di O (aminoglycosyl) 1,3 diaminocyclitols wherein the 4 O aminoglycosyl moiety 30 is defined by one of the following Formulae I and II:

H H RII-C NHT R Il CNHT 516 041 g R 1 41 R 131 2 ' 0-31 21 I 2 1,577,540 2 wherein R and R' are each hydrogen or hydroxy, R" is hydrogen or methyl, and T is hydrogen, alkyl, hydroxy-alkyl or aminoalkyl, said alkyl having up to 4 carbon atoms The invention also includes the 1-N-X-derivatives of the foregoing, wherein X is a substituent S -CH 2 Y or -C Y wherein Y is hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, hydroxyalkyl, aminoalkyl, mono or di-alkylamino-alkyl, aminohydroxyalkyl, mono or dialkylaminohydroxyalkyl, phenyl, benzyl or tolyl, said aliphatic radicals having up to 8 carbon atoms and when substituted by both amino and hydroxy groups, said groups are on different carbon atoms Additionally, when the 1,3diaminocyclitol 10 has a 5-hydroxyl, in the equatorial position, the invention embraces the 5-epi, 5deoxy, 5-epi-azido-5-deoxy and 5-epi-amino-5-deoxy derivatives thereof.

In the Formulae I and II depicted hereinabove, the stereo-chemical positions at the 6 ' carbon are indicated by wavy lines thus indicating they can be of the R or S configuration; thereby, this invention includes compounds of both 15 stereoconfigurations Compounds of this invention which are named as 2 'unsubstituted derivatives of known 4,6 di O (aminoglycosyl) 1,3 diaminocyclitols thus have the same stereoconfiguration as their 2 'substituted analogs.

More specifically, the compounds of this invention are 2 '-unsubstituted 4,6 20 di O (aminoglycosyl) 1,3 diaminocyclitols, wherein the 6 O (aminoglycosyl) is defined by the following Formula III R 2 R 3 O R 4 OH wherein R, is hydrogen or methyl, R 2 is hydrogen or hydroxymethyl, R 3 is hydrogen or hydroxy and R 4 is hydrogen, methyl or hydroxy with the proviso that 25 both R 3 and R 4 are not hydroxy.

Compounds of this invention include 2 ' unsubstituted 4,6 di O (aminoglycosyl) 1,3 diaminocyclitols wherein the substituents R, R', R" and T in the 4 O aminoglycosyl moiety of Formula I are defined as follows:

a) R and R' are hydroxy, R" is hydrogen or methyl, and T is hydrogen, e g 2 ' 30 deoxygentamicin B, 2 '-deoxygentamicin B 1, 2 '-deoxygentamicin A 3, 2 'deoxykanamycin A, and 2 '-desamino-Antibiotic JI-20 B; b) R is hydrogen, R' is hydroxy, R" is hydrogen or methyl, and T is hydrogen, e.g 2 ',3 '-dideoxygentamicin B, 2 ',3 '-dideoxygentamicin B, 2 ',3 'dideoxygentamicin A 3, 2 ',3 '-dideoxykanamycin A, and 2 '-desamino-3 'deoxy 35 Antibiotic JI-20 B; c) R and R' are hydrogen, R" and T are hydrogen or methyl, e g 2 'desaminogentamicin C,, 2 '-desaminogentamicin C,, 2 '-desaminogentamicin C 2, 2 'desaminogentamicin C 2,, 2 '-desaminogentamicin C 2 b, 2 ',3 ',4 'trideoxykanamycin A, and 2 ',3 ',4 '-trideoxygentamicin A 3 40 Other 2 '-unsubstituted derivatives of this invention are those wherein the 4 O aminoglycosyl is defined by formula II, particularly those wherein R" and T are hydrogen or methyl, such as 2 '-desaminosisomicin, 2 '-desaminoverdamicin, 2 'desamino-Antibiotic 66-40 B, 2 '-desamino-Antibiotic 66-40 D, 2 'desaminoAntibiotic G-52, 2 '-desaminoantibiotic Mu-1 and 2 '-desaminoantibiotic Mu-4 45 It is readily apparent to one skilled in the art that the compounds listed above may have multiple names, e g 2 '-deoxykanamycin A may also be named as 2 'desaminokanamycin B, 2 ',3 '-dideoxykanamycin A may also be named as 2 'desaminotobramycin, 2 '-deoxygentamicin B may also be named as 2 'desaminoAntibiotic JI-20 A, and the like 50 Preferred compounds or this invention include those wherein the 6 O aminoglycosyl is 6 O garosaminyl and the 1,3-diaminocyclitol is 2deoxystreptamine and the 5-epi or 5-deoxy derivatives thereof.

Included among the substituents contemplated for the moiety T in the novel compounds are straight and branched chain alkyl groups such as methyl, ethyl, npropyl, isopropyl, n-butyl, sec-butyl, tertiary-butyl, and isobutyl hydroxy substituted straight and branched chain alkyl groups such as /3hydroxyethyl, 1 yhydroxypropyl, /-hydroxypropyl, 8-hydroxybutyl and /-hydroxy-/p-methylpropyl; 5 amino substituted straight and branched chain alkyl groups such as p/aminoethyl, /-aminopropyl, y-aminopropyl, 8-aminobutyl, and t-methyl-v-aminopropyl.

Included among the substituents contemplated for the moiety Y in the novel compounds are straight and branched chain alkyl groups such as methyl, ethyl, npropyl, isopropyl, n-butyl, sec-butyl, tertiary-butyl, isobutyl, n-pentyl, isopentyl, / 10 methylbutyl, y-methylbutyl and /,p-dimethylpropyl: n-hexyl, 8methylpentyl, /3ethylbutyl, y-ethylbutyl, n-heptyl, isoheptyl, E-methylheptyl, /pethylpentyl, yethylpentyl, 8-ethylhexyl, y-propylbutyl, n-octyl, iso-octyl, /ethylhexyl, /ethylhexyl, E-ethylhexyl, p/-propylpentyl, y-propylpentyl: alkenyl groups such as /propenyl, /-methylpropenyl, /-butenyl, /-methyl-/p-butenyl and /-ethyl-/phexenyl; 15 cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl; cycloalkylalkyl groups such as cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl and cyclohexylmethyl; hydroxy substituted straight and branched chain alkyl groups such as E-hydroxypentyl, p/-hydroxy-ymethylbutyl, /hydroxy-o-methylpropyl, 8-hydroxybutyl, /-hydroxypropyl, y-hydroxypropyl, / 20 hydroxyethyl, cw-hydroxyoctyl; amino substituted straight and branched chain alkyl groups such as &-aminopentyl, /-aminopropyl, y-aminopropyl, 8aminobutyl, /p-amino-y-methylbutyl and w-aminooctyl; and mono-N and di-N-alkylated derivatives thereof such as the N-methyl, N,N-dimethyl, N-ethyl N,Ndiethyl, Npropyl and N,N-dipropyl, e g E-methylaminopentyl, e-dimethylaminopentyl, /3 25 methylaminopropyl, y-methylaminopropyl, /,-dimethylaminopropyl, ydimethylaminopropyl, 3-ethylaminopropyl, /-diethylaminopropyl, 8-methylaminobutyl, 8dimethylaminobutyl, /,-methylamino-y-methylbutyl, and o-methylaminobutylyaminopropyl, 8-methylaminobutyl, /P-methylamino-y-methylbutyl, and mwmethylaminobutyl; amino and hydroxy disubstituted straight and branched chain alkyl 30 groups such as /p-hydroxy-E-aminopentyl, y-hydroxy-y-methyl-8-aminobutyl,, Bhydroxy-b-aminobutyl, /p-hydroxy-y-aminopropyl, and /p-hydroxy-/p-methylyaminopropyl; and mono-N and di-N-alkylated derivatives thereof such as /hydroxy-E-methylaminopentyl, v-hydroxy-y-methyl-5-methylaminobutyl, /hydroxy-8-methylaminobutyl, /-hydroxy-S-dimethylaminobutyl, /P-hydroxy-y 35 ethylaminopropyl, /p-hydroxy-/p-methyl-y-methylaminopropyl and /,-hydroxyydimethylaminopropyl.

Particularly valuable derivatives of the compounds of this invention are the following:

a) 1-N-X-derivatives, wherein X is CH 2-Y, Y being alkyl or aminoalkyl having 40 up to 3 carbon atoms, e g 1 N ethyl 2 ',3 ' dideoxygentamicin B, and I N ethyl 2 ' desaminosisomicin; b) l-N-X-derivatives wherein X is a C-Y substituent selected from the group consisting of /-amino-ahydroxypropionyl, y 45 amino-a-hydroxybutyryl and 5-amino-a-hydroxyvaleryl, e g 1 N (/P amino a hydroxypropionyl) 2 ',3 ' dideoxygentamicin B and the stereoisomers (R, S) thereof; c) 6 '-NHT derivatives wherein T is alkyl, aminoalkyl, or hydroxyalkyl, said alkyl having up to 4 carbon atoms; and preferably hydrogen; 50 d) 5-epi or 5-deoxy analogs of those 2 '-unsubstituted compounds which have a 5-hydroxyl group in the equatorial position, e g 5 '-epi-2 'desaminosisomicin, and '-deoxy 2 '-desaminosisomicin.

Also included within this invention are the pharmaceutically acceptable acid addition salts of the 2 'unsubstituted 4,6 di O (aminoglycosyl) 1,3 55 diaminocyclitols described hereinabove, which are made according to known procedures such as by neutralizing the free base with the appropriate acid (e g.

sulfuric or hydrochloric acid) usually to about p H 4 5-5 Included among the pharmaceutically acceptable acid addition salts of this invention are those derived from organic acids such as succinic acid, fumaric acid and maleic acid, or 60 preferably, from inorganic acids such as hydrochloric, sulfuric, phosphoric and 1,577,540 hydrobromic acid The physical embodiments of the acid addition salts of this invention are characterized by being white solids which are soluble in water, sparingly soluble in most polar organic solvents and insoluble in most non-polar organic solvents.

The novel pseudotrisaccharides of this invention i e the 2 'unsubstituted 5 derivatives of 4,6 di O (aminoglycosyl) 1,3 diaminocyclitols, wherein the 4 O aminoglycosyl moiety is defined in Formulae I and 11, may be prepared by a process wherein a 6 O (aminoglycosyl) 1,3 diaminocyclitol having an unprotected 4-hydroxyl group and being suitably protected at other positions is condensed with a suitably protected glycal in the presence of an acid catalyst; any 10 protecting groups are removed by known methods and wherein, if desired, a 5-epi, 5-deoxy, 5-epi-azido-5-deoxy or 5-epi-amino-5-deoxy and/or a l-N-X and/or a 6 '-NT' derivative is prepared by known methods with X being as defined above and T' being the same as T above except for hydrogen; the last step being followed by isolating the product as such or as a pharmaceutically acceptable acid addition salt 15 (By "known methods" are meant methods hitherto used or described in the literature).

In this process the glycal is that entity referred to as the 4 O aminoglycosyl in the final compounds.

By "suitably protected" those groups are meant which are utilized to protect 20 amino and/or hydroxyl groups during chemical reactions The preferred amino protecting groups are benzyloxycarbonyl, ethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl and benzoyl and the preferred hydroxy protecting groups are acetyl and benzoyl.

The choice of the protecting group depends on various factors including 25 whether an amino or hydroxy group is being protected, subsequent reaction conditions, and conditions desired for removal The choice of the proper protecting group is within the ordinary ability of one skilled in the art.

The acid catalyst is usually selected from sulphonic acids, sulphonic acid resins, sulfuric acid, phosphoric acid, trifluoroacetic acid and sulfamic acid Most 30 preferred is p-toluenesulphonic acid.

Suitably protected starting compounds useful for producing the pseudotrisaccharides of this invention are represented by the following structural Formulae IV-IX:

NHZ NHZ HZ NHZ 3 O HHO 35 H 3 C W 3 O OW Ow Ow 1,577,540 1,577,540 Ix H 3 C H 3 C 1 ow o W wherein each Z is an amino protecting group and each W is a hydroxy protecting group Compound IV is suitably protected garamine the preparation of which is described in U K Specifications No 1,420,879 and No 1,473,178 Compounds V and VI may be obtained from Antibiotics 66-40 D and 66-40 B, respectively, by 5 applying similar reaction techniques as are described in U K Specification

1,420,879 for the conversion of sisomicin to garamine Compound VII may be obtained from kanamycin A by applying similar reaction techniques as are described in U K Specification No 1,473,178 and compounds VIII and IX may be obtained from Mu-I and Mu-4, respectively, by applying similar reaction 10 techniques as are described in U K Specification 1,420,879 Mu-l and Mu-4 are also known in the literature as mutamicin I and mutamicin 4, respectively.

The location of the functional groups in the glycal reactants will predetermine the resultant products of this invention, i e whether they are 2 'unsubstituted, 2 ',3 'di-unsubstituted, 2 ',3 ',4 '-tri-unsubstituted or 2 ',4 '-diunsubstituted pseudotri 15 saccharides For example, if the blocked monosaccharide of Example 9 ( 7) of the following Formula X:

CH 2 N 3 is reacted with the blocked disaccharide of Formula IV and the resulting blocked pseudotrisaccharide is then de-blocked, one obtains 2 ',4 'dideoxygentamicin B 20 It will be obvious that the l-N-X, and/or the 5-epi, 5-deoxy, 5-epi-azido5-deoxy and 5-epi-amino-5-deoxy derivatives of this invention may be obtained by starting with a disaccharide in which the 1,3-diaminocyclitol already contains said 1 -N-X or " 5 " derivatives before condensation with the monosaccharides or, alternatively, by introducing the foregoing groups, as well as the 6 '-N-T' group, into the 25 corresponding 2 '-unsubstituted pseudo-trisaccharides of this invention.

Derivatization of the pseudotrisaccharides of this invention may be accomplished by various means The l-N-alkyl and l-N-acyl derivatives of the compounds of this invention may be prepared according to procedures set forth in U K Specification No 1,473,733 Utilizing one of the methods described therein, 30 reaction of an acid addition salt of a 2 ' unsubstituted 4,6 di O aminoglycosyl 1,3 diaminocyclitol (e g 2 ' desaminosisomicin sulfate) in an inert protic solvent containing water with one equivalent of a hydride donor reducing agent (e g cyanoborohydride) and at least one equivalent of an aldehyde (e g acetaldehyde) yields a I N alkyl 2 ' unsubstituted derivative of this 35 invention (e g I N ethyl 2 ' desaminosisomicin) Similarly, by utilizing one of the methods described in said U K Specification l-N-acyl derivatives may be prepared Reaction of an acid addition salt of a 2 ' unsubstituted 4,6 di O (aminoglycosyl) 1,3 diaminocyclitol (e g 2 '-desaminosisomicin sulfate) in an inert protic solvent containing water, with one equivalent of an organic or 40 inorganic base (e g triethylamine), followed by the addition of a molar excess of an acylating agent (e g acetic anhydride) yields a I-N alkanoyl 2 ' unsubstituted derivative of this invention (e g I N acetyl 2 ' desaminosisomicin).

The 5-epi, 5-epi-azido-5-deoxy, 5-epi-amino-5-deoxy derivatives of 4,6 di O (aminoglycosyl) 1,3 diamino cyclitols may be prepared according to 45 procedures similar to those described in U S Patents No 4,000,261 and No.

4,000,262 Thus the 5-epi derivatives of the compounds of this invention may be prepared by the reaction of a 5 O hydrocarbonsulfonyl 2 ' unsubstituted aminoglycoside (e g 5 O methanesulfonyl 2 ' desaminosisomicin) having other hydroxyl and amino groups suitably protected, with dimethylformamide at elevated temperatures, followed by removal of the protecting groups to form the 5 epi 2 ' unsubstituted aminoglycoside (e g 5 epi 2 ' desaminosisomicin).

The 5 epi azido 5 deoxy and 5 epi amino 5 deoxy derivatives of the compounds of this invention may be prepared from a 5 O hydrocarbonsulfonyl 2 ' unsubstituted aminoglycoside (e g 5 O 10 methanesulfonyl 2 ' desaminosisomicin) having all other amino and hydroxyl groups suitably protected, by reaction thereof with an alkali metal azide in an organic solvent followed by the reaction of the resulting 5 epi azido 5 deoxy 2 ' unsubstituted protected aminoglycoside (e g 5 epi azido 5 deoxy protected 2 ' desaminosisomicin) with base to remove the protecting 15 groups Alternatively, reaction of an O and N-protected 5 epi azido 5 deoxy 2 ' unsubstituted derivative with hydrogen in the presence of a catalyst or with an alkali metal in liquid ammonia, and thence cleavage of any remaining hydroxyl or amino protecting groups forms a 5 epi amino 5 deoxy 2 ' unsubstituted derivative (e g 5 epi amino 5 deoxy 2 ' 20 desaminosisomicin).

The 5-deoxy derivatives of this invention may be prepared according to procedures similar to those described in Belgium Patent 844,551 (X=hydrogen in Formula I of that patent) For instance, a 5 O thioformyl 2 ' unsubstituted aminoglycoside (e g 5 O thioformyl 2 ' desaminosisomicin) having all amino 25 functions and all primary and secondary hydroxyl groups protected, is reacted with an organotin hydride (preferably tri-n-butyl tin) in an inert protic solvent under an inert atmosphere at temperatures of about 100 C, followed by removal of the protecting groups to obtain the 5-deoxy-2 '-unsubstituted aminoglycoside (e g 5 deoxy 2 ' desaminosisomicin) 30 The 6 '-N-alkyl derivatives of this invention may be prepared according to known methods or by procedures similar to those described in U K Specification

No 1,473,733 for alkylating the 1-amino group Said 6 '-N-alkyl derivatives may be prepared from a 2 ' unsubstituted 6 ' N unsubstituted aminoglycoside (e g 2 'desaminosisomicin) having protecting groups on all other amino functions, by 35 reaction with an aldehyde or a ketone (e g acetaldehyde), followed by reaction in situ of the 6 '-N-substituted intermediate thereby formed with a hydride reducing agent, and then removal of any N-protecting groups to obtain the 6 ' N alkyl 2 ' unsubstituted aminoglycosides (e g 6 ' N ethyl 2 ' desaminosisomicin).

The invention described herein above is illustrated in detail herein below in 40 Examples 1-8 Starting compounds useful in the preparation of the compounds of this invention may be prepared as illustrated in Example 9.

EXAMPLE I

2 '-Unsubstituted-4,6-di-O-(Aminoglycosyl)-l,3Diamino-cyclitols 45 A 2 '-Deoxygentamicin B Dissolve 1 gm of 1,3,3 ' tri N benzyloxycarbonyl 2 ' O acetylgaramine, 0 7 gm of 6 acetamido 6 deoxy 3,4 di O benzyl D glucal, and 5 mg of p-toluenesulphonic acid monohydrate in 10 ml anhydrous benzene and heat the solution at 50 C for 5 hours Cool the solution and wash with 50 dilute aqueous sodium bicarbonate and dry over Mg SO 4 Concentrate the solution in vacuo to a residue and chromatograph on a silica gel column eluating with 1/,, methanol in chloroform Follow the eluates by thin layer chromatography to obtain a mixture containing 1,3,3 " tri N benzyloxycarbonyl 3 ',4 ' di Obenzyl 6 ' N acetyl 2 " O acetyl 2 ' deoxygentamicin B Concentrate 55 the eluates to a residue and dissolve said residue in 5 ml of tetrahydrofuran and 50 ml distilled liquid ammonia To the solution add, in small pieces, 0 7 gm of sodium, after 20 minutes allow the solvent to evaporate Add 5 ml water and 0 5 gin sodium hydroxide to the residue and reflux the resulting solution for 3 hours.

Cool the solution and stir with an excess of Amberlite (Registered Trade Mark), 60 IRC-50 (H ) ion exchange resin, filter the resin and wash with water Elute the resin with 3 , aqueous ammonium hydroxide, concentrate the eluates in vacuo and lyophilize Chromatograph the residue on a silica gel column using the lower phase of a chloroform-methanol-15 /0 ammonium hydroxide ( 2:1:1) as the eluant Follow 1,577,540 the eluates by thin layer chromatography and combine like eluates containing 2 'deoxygentamicin B Concentrate the combined eluates, dissolve the residue in water and pass over Amberlite IRA-401 S (OH ) resin and lyophilize to obtain 2 'deoxygentamicin B, lal 26 + 136 7 (H 20); 8 (D 20) 1 18 ( 3 H, s, C-CH 3), 2 57 ( 3 H, s, N-CH 3), 5 03 (IH, d, J 4 Hz, H 1 ") and 5 47 ( 1 H, broad d, J 3 5 Hz, H) ; cd (TA Cu) 5 l O l 288 -6,360.

B Other 2 '-Unsubstituted Aminoglycosides In a manner similar to the foregoing, react the following appropriately blocked glucals and disaccharides.

1 6 -(R) -6 -azido -6 -deoxy 6 C methyl -3,4 -di -O -benzyl 10 D glucal and 5,2 ',4 ' tri O acetyl 1,3,3 ' tri N benzyloxycarbonylgaramine, 2.6 acetamido 6 deoxy 3,4 di O benzyl D -glucal and O ( 3 amino 3 deoxy 3 N methyl 3 N benzyloxycarbonyl 2,4 di O acetyl /3L arabinopyranosyl) ( 1-,6) 1,3 di N benzyloxycarbonyl 15 O acetyl 2 deoxy D streptamine, 3 6 acetamido 6 deoxy 3,4 di O benzyl D glucal and O ( 3 amino 3 deoxy 3 N benzyloxycarbonyl 2,4,6 tri O acetyl a D glucopyranosyl) ( 16) 1,3 di N benzyloxycarbonyl 5 O acetyl 2 deoxy D streptamine, 20 4 6 (S) 6 azido 6 -deoxy -6 C methyl -3,4 di O benzyl D glucal and 5,2 ',4 ' tri O acetyl 1,3,3 ' tri Nbenzyloxycarhonylgaramine, to obtain respectively, the following 2 '-unsubstituted aminoglycosides:

1 2 '-deoxygentamicin B 1, 25 2 2 '-deoxygentamicin A 3, 3 2 '-deoxykanamycin A, and 4 2 '-desamino-Antibiotic JI-20 B. EXAMPLE 2

2 ',3 '-di-Unsubstituted-4,6-di-O-(aminoglycosyl) 30 1,3-Diaminocyclitols A 2 ',3 '-Dideoxygentamicin B Dissolve 19 52 gm of 5,2 ',4 '-tri-O-acetyl 1,3,3 ' tri N benzyloxycarbonylgaramine, 7 1 gm of 6 azido 3,6 dideoxy 4 O acetyl D glucal and 180 mg of p-toluene-sulfonic acid monohydrate in 200 ml anhydrous benzene Heat 35 the solution at 45 C for 5 hours and let it adjust to room temperature overnight.

Wash the solution with dilute aqueous sodium bicarbonate solution Dry the benzene solution over Mg SO 4 and then concentrate in vacuo to a residue.

Chromatograph the residue on a silica gel column eluting with 0 25 o methanol in chloroform Follow the eluates by thin layer chromatography and concentrate the 40 less polar eluates to a residue Dissolve 5 gm of the residue in 100 ml of 20 % aqueous dioxane, and hydrogenate for 16 hours at room temperature and 4 atmospheres in the presence of 500 mg of 10 O Pd/C Filter off the catalyst and evaporate the filtrate to dryness Heat the residue at reflux in a solution of 3 gms of potassium hydroxide in 20 ml distilled water for 18 hours Cool the solution and stir 45 with Amberlite IRC-50 (H") ion exchange resin, filter the resin and wash with water Elute the resin with 30 aqueous ammonium hydroxide, concentrate the combined eluates in vacuo and chromatograph the residue on a 50 gm silica gel column using the lower phase of a chloroform-methanol-150,% aqueous ammonium hydroxide ( 2:1:1) as the eluant Follow the eluates by thin layer chromatography, 50 combine like eluates containing the desired product and concentrate the combined eluates, dissolve the residue in water and pass over Amberlite IRA-4015 (OH') resin and lyophilize to obtain 2 ',3 '-dideoxygentamicin B, lal 6 + 171 (H 20); 8 (D 20) 1.15 ( 3 H, s, C-CH 3); 2 46 ( 3 H, s, N-CH 3), 3 75 (IH, dd, J= 4, 11 Hz, H-2 "); 3 96 (IH, d, J= 12 5 Hz, H-5 " eq), 5 02 (IH, d, J= 4 Hz, H-I"), and 5 27 ppm (IH, 55 broad s, W 1/2 = 5 Hz, H-I'); cd (TA Cu) l 01288 -10,100.

B Other 2 ',3 '-Di-Unsubstituted Aminoglycosides In a manner similar to the foregoing, react the following appropriately blocked glucals and disaccharides, 1 6 (R) 6 azido 3,6 dideoxy -6 C methyl 4 O benzyl D 60 glucal and 5,2 ',4 ' tri O acetyl 1,3,3 ' tri N benzyloxycarbonylgaramine, 1.577-540 2.6 azido 3,6 dideoxy -4 O acetyl D glucal and O ( 3 amino 3 deoxy 3 N methyl 3 N benzyloxycarbonyl 2,4 di O acetyl /p L arabinopyranosyl) ( 1- 6)1,3 di N benzyloxycarbonyl 5 O acetyl 2 deoxy D streptamine.

3 6 azido 3,6 dideoxy -4 O acetyl D glucal and O ( 3 -amino 5 3 deoxy 3 N benzyloxycarbonyl 2,4,6 tri O acetyl a D glucopyranosyl) ( 1-,6) 1,3 di N benzyloxycarbonyl 5 O acetyl 2 deoxy D streptamine, 4 6 (S) 6 -azido 3,6 dideoxy -6 C methyl 4 O benzyl -D glucal and 5,2 ',4 'triO acetyl 1,3,3 'tri N benzyloxycarbonylgaramine, 10 to obtain, respectively, the following 2 ',3 '-di-unsubstituted aminoglycosides:

1 2 ',3 '-dideoxygentamicin B, 2 2 ',3 '-dideoxygentamicin A 3, 3 2 ',3 '-dideoxykanamycin A, and 4 2 '-desamino-3 '-deoxy-Antibiotic JI-20 B 15 EXAMPLE 3

2 ',3 ',4 '-Tri-Unsubstituted 4,6-di-O-(aminoglycosyl)1,3-Diaminocyclitols A 2 '-Desaminogentamicin C 1 a Dissolve 4 33 gm of 5,2 ',4 ' tri O acetyl 1,3,3 ' tri N 20 benzyloxycarbonylgaramine, 1 54 gm of 6 azidomethyl 5,6 dihydropyran and mg of p-toluenesulfonic acid monohydrate in 45 ml of benzene and heat the solution at 40 C overnight Cool the solution and wash with dilute aqueous sodium bicarbonate and dry over Mg SO 4 Concentrate the solution in vacuo to a residue and chromatograph on a silica gel column using 0 25 % methanol in chloroform as 25 the eluant Follow the eluates by thin layer chromatography, combine the less polar eluates and evaporate Dissolve the residue in 20 ml 50,, aqueous dioxane, and add 200 mg of 5 % Pd/C and 5 ml of O IN HCI and hydrogenate at room temperature at 4 atmospheres for 16 hours Filter, and evaporate the filtrate in vacuo Dissolve the residue in 4 ml of 20 aqueous potassium hydroxide and heat 30 at reflux under argon for 16 hours Cool the solution and adjust the p H to 11 by addition of dilute sulfuric acid Concentrate the solution to a small volume and add ml absolute ethanol dropwise while stirring Filter the resultant mixture and reduce the filtrate to dryness.

Dissolve the residue in the lower phase of a chloroform methanol-10 35 ammonium hydroxide ( 2:1:1) solvent system and chromatograph on a silica gel column eluting with the same solvent Monitor the eluates by thin layer chromatography Evaporate the less polar eluates containing 2 '-desaminogentamicin Ca Dissolve the residue in water and pass over Amberlite IRA4015 (OH ) resin and lyophilize to obtain 2 '-desaminogentamicin Ca, lal 26 + 166 (H 20); 40 8 (D 20) 1 17 ( 3 H, s, C-CH 3), 2 48 ( 3 H, s, N-CH 3), 3 71 (IH, dd, J= 4, 11 Hz, H2 "), 4 0 (IH, d, J= 12 5 Hz, H-5 " eq), 5 04, (IH, d, J= 4 Hz, H-I") and 5 30 (IH, broad s, W 1/2 = 5 Hz, H-I'); cd (TA Cu) l 01259 -7,760.

B Other 2 ',3 ',4 '-Tri-Unsubstituted Aminoglycosides In a manner similar to the foregoing react the following appropriately blocked 45 glucals and disaccharides, 1 6 (R) 6 amino 3,4,6 trideoxy 6 C methyl 6 N methyl 6 N benzyloxycarbonyl D glucal and 5,2 ',4 'triO acetyl 1,3,3 'tri N benzyloxycarbonylgaramine, 2 6 -(R) -6 amino 3,4,6 -trideoxy -6 C methyl -6 -N benzyloxy 50 carbonyl D glucal and 5,2 ',4 ' tri O acetyl 1,3,3 ' tri Nbenzyloxycarbonylgaramine, 3 6 (S) 6 amino 3,4,6 trideoxy 6 C methyl 6 Nbenzyloxycarbonyl D glucal and 5,2 ',4 'tri O acetyl 1,3,3 ' tri N benzyloxycarbonylgaramine, 554 6 amino 6 N methyl 6 N benzyloxycarbonyl 3,4,6 trideoxy D glucal and 5,2 ',4 ' tri O acetyl 1,3,3 ' tri N benzyloxycarbonylgaramine, 6 azidomethyl 5,6 dihydropyran and O ( 3 amino 3 deoxy 3 N benzyloxycarbonyl 2,4,6 tri O acetyl a D glucopyranosyl) 60 ( 1-o 6)1,3 di N benzyloxycarbonyl 5 O acetyl 2 deoxy D streptamine, 1,577,540 À 7-w | 6.6 azidomethyl 5,6 dihydropyran and O ( 3 amino 3 deoxy 3 N methyl 3 N benzyloxycarbonyl 2,4 di O acetyl A Larabinopyranosyl) ( 16) 1,3 di N benzyloxycarbonyl 5 O acetyl 2 deoxy D streptamine; to obtain, respectively, the following 2 ',3 ',4 'triunsubstituted aminoglycosides: 5 1 2 '-desaminogentamicin C, 2 2 '-desaminogentamicin C 2, 3 2 '-desaminogentamicin C 2 a, 4 2 '-desaminogentamicin C 2 b, 5 2 ',3 ',4 '-trideoxykanamycin A, and 6 2 ',3 ',4 '-trideoxygentamicin A 3 10 EXAMPLE 4

2 '-Unsubstituted-4,6-di-O-(Aminoglycosyl) 1,3-diaminocyclitols having 4 ',5 '-Unsaturation A 2 '-Desaminosisomicin 15 Dissolve 410 mg of 5,2 ',4 'tri O acetyl 1,3,3 'tri N benzyloxycarbonylgaramine, 150 mg of 6 azido 3,6 dideoxy 4 O acetyl D glucal and 3 8 mg of p-toluenesulfonic acid monohydrate in 35 ml anhydrous benzene Heat the solution at 45 C for 5 hours and let it adjust to room temperature overnight Wash the benzene solution with dilute aqueous sodium 20 bicarbonate solution, dry the benzene solution over Mg SO 4 and concentrate in vacuo to a residue Dissolve the residue in 250 ml dichloromethane and photolyse with a 450 W Hg lamp for 7 hours Evaporate the solvent Repeat the reaction 19 times Combine the photolysis product of the 19 runs and dissolve in 170 ml 95 %/ ethanol Add 6 8 ml of triethylamine and stir at room temperature for 5 hours 25 Evaporate the solvent and chromatograph the residue on silica gel using chloroform as the eluant to obtain the 4 ',5 '-unsaturated aldehyde.

Dissolve 1 62 gm of the 4 ',5 '-unsaturated aldehyde in 8 ml of tetrahydrofuran.

Add this to a solution of 9 6 gm ammonium acetate in 45 ml methanol Stir for 30 minutes and then add 960 mg sodium cyanoborohydride and stir at room 30 temperature for 2 5 hours and overnight at 4 C Add 150 ml water, extract with 3 x 100 ml chloroform, dry the chloroform extracts over Mg SO 4 and evaporate in vacuo Dissolve the residue in 15 ml of dimethylsulphoxide, add 2 6 gm of potassium hydroxide in 5 ml of water and stir at room temperature for 48 hours.

Adjust the p H of the reaction mixture to 11 by adding dilute H Cl, then stir the 35 solution with IRC-50 (H ) ion exchange resin Filter the resin, wash with water and then elute with 60, aqueous ammonium hydroxide Evaporate the ammoniacal eluate to dryness and chromatograph the residue on silica gel in the lower phase of a chloroform-methanol-7 y ammonium hydroxide ( 2:1:1) system Follow the eluates by thin layer chromatography, combine like fractions containing 2 ' 40 desaminosisomicin, concentrate, pass over Amberlite IRA-401 S (OH ) resin and lyophilize to obtain 2 '-desaminosisomicin, lal 26 + 184 3 (H 20); 8 (D 20) 1 18 ( 3 H, s, C-CH 3), 2 49 ( 3 H, s, N-CH 3), 3 76 (IH, dd, J= 4, 11 Hz, H-2 "), 4 03 (IH, d, J= 12 5 Hz, H-5 " eq), 4 92 (IH, m, H-4 '), 5 05 (IH, d, J= 4 Hz, H-I") and 5 46 (IH, distorted triplet, W 1/2 = 6 Hz, H-I') 45 B Other 2 '-Unsubstituted Aminoglycosides having a 4 ',5 '-Unsaturation In a manner similar to the foregoing, react the following appropriately blocked glucals and disaccharides, 1 6 (S) -6 azido -3,6 dideoxy -6 C methyl -4 O benzyl D glucal and 5,2 ',4 'triO acetyl 1,3,3 'tri N benzyloxycarbonylgaramine, 50 2.6 azido 3,6 dideoxy 4 O acetyl D glucal and O ( 3 amino 3 deoxy 3 N methyl 3 N benzyloxycarbonyl 2,4 di O acetyl a D xylopyranosyl) ( 1-6) 1,3 di N benzyloxycarbonyl 5 O acetyl 2 deoxy D streptamine, 3 6 azido 3,6 dideoxy 4 O acetyl D glucal and O ( 3 amino 55 3 deoxy 3 N methyl 3 N benzyloxycarbonyl 2,4 di O acetyl /3 L arabinopyranosyl) ( 1-6) 1,3 di N benzyloxycarbonyl 5 O acetyl 2 deoxy D streptamine, 4 6 azido 3,6 dideoxy 4 O acetyl D glucal and 5,2 ',4 'triO acetyl 1,3,3 ' tri N benzyloxycarbonylgaramine, (for this preparation 60 methylamine hydrochloride is used instead of ammonium acetate in the above procedure), 1.577 540 5.6 azido 3,6 dideoxy -4 O acetyl D glucal and O -( 3 amino 3 deoxy 3 N methyl 3 N benzyloxycarbonyl 4 C methyl 24 di O acetyl /3 L arabinopyranosyl ( 1-6) 1,3 di N benzyloxycarbonyl 2,5 di O acetyl D streptamine, 6 6 azido 3,6 dideoxy 4 O acetyl D -glucal and O ( 3 amino 5 3 -deoxy 3 N methyl 3 N benzyloxycarbonyl 4 C methyl 2,4 di O acetyl /3 L arabinopyranosyl) ( 1-6) 1,3 di N benzyloxycarbonyl 2,5 di O acetyl 2 epi D streptamine, to obtain, respectively, the following 2 '-unsubstituted aminoglycosides having 4 ', 5 'unsaturation: 10 1 2 '-desaminoverdamicin, 2 2 '-desamino-Antibiotic 66-40 B, 3 2 '-desamino-Antibiotic 66-40 D, 4 2 '-desamino-Antibiotic G-52, 5 2 '-desaminomutamicin 1, and 15 6 2 '-desaminomutamicin 4.

EXAMPLE 5 l-N-X-Derivatives of 2 '-Unsubstituted-4,6-di-O-(Aminoglycosyl) I,3-Diaminocyclitols A I-N-Ethyl-2 ',3 '-Dideoxygentamicin B 20 Dissolve 2 77 gm of 4 O acetyl 6 azido 3,6 dideoxy D glucal, 6.14 gin5,2 ',4 'tri O acetyl 1,3,3 'tri N benzyloxycarbonyl I N ethylgaramine and 50 mg p-toluene-sulphonic acid in 100 ml anhydrous benzene and stir at 40 C for 18 hours Cool the solution and wash with dilute Na HCO 3, dry over Mg SO 4 and concentrate in vacuo Chromatograph on silica gel in O 25 / 25 methanol in chloroform and follow the eluates by thin layer chromatography.

Dissolve the product ( 2 56 gm) from the combined less polar fractions in 125 ml.

% aqueous dioxane, add 10 pd/C and hydrogenate at 4 atmospheres for 16 hours Filter the reaction mixture and evaporate the filtrate in vacuo to a residue.

Heat the residue in 20 ml 2 N sodium hydroxide at 100 C for 16 hours under argon 30 Cool the solution and adjust the p H to 11 with the addition of IN H 25 04.

Concentrate the solution to a small volume and add 200 ml absolute ethanol dropwise while stirring Filter the mixture and reduce the filtrate to dryness Take up the residue in the lower phase of a chloroform-methanol-10 ammonium hydroxide ( 2:1:1) solvent system and chromatograph on a silica gel column in the 35 same solvent Monitor the eluates by thin layer chromatography Evaporate the combined eluates containing the desired product and dissolve the residue in water, pass over Amberlite IRA 401 S (OH ) resin and lyophilize to obtain l-Nethyl-2 ',3 'dideoxygentamicin B, lal"+ 137 5 (Me OH); 8 (D 20) 1 05 ( 3 H, t, J= 6 5 Hz, CH 2CH 3), 1 19 ( 3 H, s, C-CH 3), 2 51 ( 3 H, s, N-CH 3), 3 76 (IH, dd, J= 4, 11 Hz, H-2 "), 40 4.0 (IH, d, J= 12 5, H-5 " eq), 4 96 ( 1 H, d, J= 4 Hz, H-I") and 5 32 (IH, broad s, Wl/2 = 6 Hz, H,); cd (TA Cu) l O l 287 -8,300.

B Other l-N-X-derivatives of 2 ',3 '-dideoxygentamicin B In the above Example, by utilizing as starting compound the l-N-propyl or 1N-8-aminobutyl analog of 5,2 ',4 'tri O acetyl 1,3,3 'tri N benzyloxy 45 carbonylgaramine, there is obtained the I N propyl 2 ',3 ' dideoxygentamicin B and I N ( 8 aminobutyl) 2 ',3 ' dideoxygentamicin B, respectively.

C I N (S -/3 amino a hydroxypropionyl) 2 ',3 'dideoxygentamicin B Dissolve 0 75 gm of 2 ',3 '-dideoxygentamicin B in 70 ml of aqueous ( 2 5, ) dimethyl sulphoxide and to this solution add, at room temperature, 88 8 mg 50 hydrated cupric diacetate and 111 mg nickel (II) diacetate Stir the reaction for 10 minutes and add 132 2 mg of N-benzyloxycarbonyloxysuccinimide and stir for 2 hours Then triturate the reaction mixture with dry ether and filter off the solid.

Dissolve the solid in 50 ml methanol-ammonium hydroxide ( 9:1) and bubble H 25 through the solution for 3 minutes Filter the resulting mixture through Celite 55 (Registered Trade Mark) and stir the filtrate with Amberlite IRA 401 S (OH ) resin until the solution becomes colorless Filter, evaporate the filtrate and dissolve the residue in 25 ml of aqueous ( 50 %) methanol To this mixture add portionwise 1 68 gm of the N-hydroxysuccinimidyl acid ester of N benzyloxy carbonyl L isoserine Stir the reaction for 2 hours and then partition the reaction mixture 60 1,577,540 between water and chloroform Wash the chloroform layer with water, dry over Mg SO 4 and then evaporate to dryness Chromatograph the resultant residue on silica gel eluting with chloroform-methanol-ammonium hydroxide ( 28 % NH 3) ( 12:10:0 5) to obtain I N (S / benzyloxycarbonylamino a hydroxypropionyl) 3,6 ' di N benzyloxycarbonyl 2 ',3 'dideoxygentamicin 5 B Hydrogenate 0 35 gm of said compound in 20 ml of aqueous ( 30 %) dioxane at one atmosphere with 25 mg of 5 % Pd/C When the hydrogenation is complete filter the reaction and reduce the filtrate to a residue Chromatograph the residue on silica gel in the lower phase of a chloroform-methanol-ammonium hydroxide ( 28 % NH 3) ( 2:1:1) solvent system Follow the eluates by thin layer chromatography, combine 10 like eluates containing the desired product Concentrate the eluates, dissolve the residue in water and pass over Amberlite IR 401 S (OH@) resin and lyophilize to obtain I N (S /3 amino a hydroxypropionyl) 2 ',3 'dideoxygentamicin B, lal 216 + 24 5 (Et OH); S(D 20) 1 13 ( 3 H, s, C-CH 3); 2 43 ( 3 H, s, N-CH 3), 2 47 (IH, d, J= 11 Hz, H-3 "), 4 05 ( 1 H, d, J= 12 5 Hz, H-5 " eq), 4 10 (IH, t, J= 5 Hz, 15 H-2 '), 5 05 (IH, d, J= 4 Hz, H-I") and 5 28 (IH, broad s, W 1/2 = 5 0 Hz, H-I').

D Other l-N-X-derivatives of 2 ',3 '-dideoxygentamicin B In the above Example if the N-hydroxysuccinimidyl acid ester of S 4 benzyloxycarbonylamino 2 hydroxybutyric acid or S 5 benzyloxycarbonylamino 2 hydroxyvaleric acid is substituted for N 20 benzyloxycarbonyl L isoserine there is obtained respectively 1 N (S y amino a hydroxybutyryl) 2 ',3 'dideoxygentamicin B or I N (S 8 amino a hydroxyvaleryl) 2 ',3 ' dideoxygentamicin B. E I N (S /3 amino a hydroxypropionyl) 2 ' desaminosisomicin In the above Example 5 C if 2 '-desaminosisomicin is substituted for 2 ', 3 ' 25 dideoxygentamicin B and by following substantially the procedure of this Example there is obtained 1N (S / amino a hydroxypropionyl) 2 ' desaminosisomicin, 8 (D 20) 1 17 (s, 3, 4 "-CH 3); 2 42 (s, 3, NCH 3): 2 44 (d, J 2,,3,=ll Hz 1.

H-3 "); 3 33 (d, J 5,,ax 5,,eq= 12 Hz, 1, H 5 ",j) 3 67 (dd, J 2,,3,,=ll Hz; J 2,,,,= 4 Hz, 1, H-2 "); 4 10 (d, J 5,,,,85,,x= 12 Hz, 1, H-5 eq); 4 14 (t, J 2,,,3,,,= 6 Hz, 1, 30 H-2 "') 4 89 (min, 1, H-4) 5 05 (d, J,,2,,= 4 Hz, 1, H-l") and 5 46 (m, 1, H-I').

EXAMPLE 6

5-Deoxy-Derivatives of 2 '-Unsubstituted-4,6-di-O-(aminoglycosyl) 1,3-diaminocyclitols A 5-Deoxy-2 '-Desaminosisomicin 35 1 1,3,6 'Tri N Ethoxycarbonyl 5 O Thioformyl 2 " O Benzoyl 3 ",4 " N,O Carbonyl 2 ' desaminosisomicin To a stirred solution of phosgene (ca 4 gms) in benzene ( 30 ml) add dimethylformamide ( 3 gms) Stir the resultant suspension for 30 minutes, then evaporate, add dichloromethane ( 30 ml) to the resultant residue and cool in an ice 40 bath To the cooled solution add 1,3,6 ' tri N ethoxycarbonyl 2 " O benzoyl 3 ",4 " N,0 carbonyl 2 ' desaminosisomicin ( 6 9 gms) in dichloromethane ( 70 ml) and pyridine ( 7 ml) Stir the reaction mixture in an ice bath for 15 minutes, the bubble dry hydrogen sulfide gas through the reaction mixture for 5 minutes Wash the solution with water, then dilute sulfuric acid and 45 finally with sodium bicarbonate Dry the solution over sodium sulfate, filter and evaporate Dissolve the resultant residue in methylene chloride and pour onto a silica gel column ( 100 gms) Elute first with chloroform, then elute with 2 methanol in chloroform combining like eluates containing the desired compound as determined by thin layer chromatography, evaporate the combined eluates in 50 vacuo, dissolve the resultant residue in chloroform ( 30 ml) and add the chloroform solution to hexane ( 500 ml) with vigorous stirring Separate the resultant precipitate by filtration, then dry the precipitate at 60 C at I millimeter pressure to obtain 1,3,6 'tri N ethoxycarbonyl 5 O thioformyl 2 " O benzoyl 3 ",4 "N,O carbonyl 2 ' desaminosisomicin 55 2 1,3,6 'Tri N Ethoxycarbonyl 2 " O Benzoyl 3 ",4 "N,O carbonyl deoxy 2 ' Desaminosisomicin Dissolve 1,3,6 'tri N ethoxycarbonyl 5 O thioformyl 2 " O benzoyl 3 ",4 " N,O carbonyl 2 ' desaminosisomicin ( 5 0 gms) and tri-nbutylstannane ( 3 5 gins) in dry toluene ( 90 ml) and heat at reflux temperature for 60 II 1,577,540 12 1,577,540 12 hours under an atmosphere of argon Evaporate the reaction mixture in vacuo, then wash the resultant residue twice by decantation with hexane and dissolve in chloroform Apply the chloroform solution to a column of silica gel ( 6 x 30 cm) containing from 2 to 3 W calcium hydroxide Elute the column with chloroform to remove the tin compounds, then develop the column with a 30 ethanolchloroform 5 solvent mixture, monitoring the fractions by thin layer chromatography (in 5/,, methanol:chloroform) Combine those fractions containing 1,3,6 ' tri Nethoxycarbonyl 2 " O benzoyl 3 ",4 " N,O carbonyl 5 deoxy 2 ' desaminosisomicin, evaporate the combined fractions and dry the resultant residue at 60 C in vacuo to obtain 1,3,6 'tri N ethoxycarbonyl 2 " O benzoyl 10 3 ",4 "N,O carbonyl 5 deoxy 2 ' desaminosisomicin.

3 5-Deoxy-2 '-desaminosisomicin To a solution of 1,3,6 'triN ethoxycarbonyl 2 " O benzoyl 3 ",4 " N,0 carbonyl 5 deoxy 2 ' desaminosisomicin ( 2 1 gms) in dimethyl sulfoxide ( 18 ml) add potassium hydroxide ( 4 gins) in water ( 6 ml) and stir under 15 an atmosphere of argon for two days Add water ( 50 ml) to the reaction mixture, stir the solution with IRC-50 (H ) resin until all the aminoglycoside has been absorbed onto the resin, pour the resin onto the column, then elute with 2 N ammonium hydroxide Combine the like eluates containing the desired product as determined by thin layer chromatography, evaporate the combined eluates, and 20 chromatograph the resultant residue on silica gel ( 50 gms) eluting with the lower phase of a chloroform:methanol:concentrated ammonium hydroxide ( 2:1:1) system Combine the like fractions containing the desired product as determined by thin layer chromatography, evaporate the combined fractions, add water to the resultant residue and pass over a small quantity of IRA-401 S (OH ) resin 25 Lyophilize the eluate to obtain 5 deoxy 2 ' desaminosisomicin.

EXAMPLE 7

5-Epi-Derivatives of 2 '-unsubstituted 4,6-di-O-(aminoglycosyl) 1,3-diaminocyclitols A 5-Epi-2 'Desaminosisomicin 30 1 1,3,6 ' tri N Benzyloxycarbonyl 5 O Methanesulfonyl 2 " O Benzoyl 3 ",4 " N,O Carbonyl 2 ' desaminosisomicin Dissolve 2 5 gms of 1,3,6 ' tri N benzyloxycarbonyl 2 " O Benzoyl 3,4 " N,O Carbonyl 2 ' desaminosisomicin in 15 ml of dry pyridine Cool the solution to 10 C and add 4 ml of methanesulfonyl chloride over a period of 10 35 minutes, allow the reaction mixture to stand overnight, then concentrate the reaction mixture under vacuum at 25 C Extract the residue with 150 ml of acidfree chloroform Wash the chloroform extracts with water and dry over sodium sulfate Evaporate the chloroform to give 1,3,6 ' tri N benzyloxycarbonyl 5 O methanesulfonyl 2 ", O benzoyl 3 ",4 " N,0 carbonyl 2 ' 40 desaminosisomicin.

2 5-Epi-2 "-Desaminosisomicin Add 2 gms of 1,3,6 ' tri N benzyloxycarbonyl 5 O methanesulfonyl 2 " O benzoyl 3 ",4 "N,O carbonyl 2 ' desaminosisomnicin to 15 ml of dimethylformamide, heat at reflux for 18 hours and then evaporate the 45 solution to a residue Dissolve the residue in a mixture of 10 ml of tetrahydrofuran and 50 ml of liquid ammonia Slowly add 2 gms of sodium to the stirred mixture and continue to stir for 2 hours Allow the ammonia to evaporate by warming to room temperature overnight Dissolve the resultant residue in 10 ml of 5 sodium hydroxide and heat at 100 C for 4 hours Cool and pass the solution through IRC 50 (H ) resin Wash the resin well with water and elute the product with 100 ml of IN ammonium hydroxide Concentrate the ammonium hydroxide eluate to a residue comprising 5-epi-2 '-desaminosisomicin Purify the product by chromatography on a silica gel column eluting with the lower phase of a chloroform:methanol:15 W ammonium hydroxide ( 2:1:1) solvent system Combine 55 the like eluates as determined by thin layer chromatography and lyophilize to a residue to obtain 5-epi-2 '-desamino-sisomicin.

B By substituting 1,3,6 ' tri N benzyloxycarbonyl 4 ',2 " di O benzoyl 3 ",4 " N,O carbonyl 2 ',3 ' dideoxygentamnicin B for 1,3,6 ' tri N benzyloxycarbonyl 2 " O benzoyl 3 ",4 " N,0 carbonyl 2 ' 60 desaminosisomicin in step Al and by following the procedure of this Example, there is obtained 5-epi-2 ',3 '-dideoxygentamicin B; Mass Spectrum (M:1)+ 451; lal 26 + 143 (H 20).

EXAMPLE 8

5-Epi-Azido-5-deoxy and 5-Epi-Amino-5-deoxy Derivatives of 2 '-Unsubstituted-4,6-di-O-(aminoglycosyl) 5 1,3-diaminocyclitols A 5-Epi-Azido-5-deoxy-2 '-desaminosisomicin 1 1,3,6 'tri N Benzyloxycarbonyl 5 Epi Azido 5 Deoxy 2 "O Benzoyl 3 ",4 " N,O Carbonyl 2 ' Desaminosisomicin Dissolve 2 gm of the product of Example 7 A 1 I in 15 ml of dry 10 dimethylformamide Stir the mixture and add 1 5 gm of sodium azide Keep the reaction mixture under argon at 120 C overnight Concentrate the solution under high vacuum Extract the residue with 200 ml of acid-free chloroform Wash the chloroform extracts with water and dry over sodium sulfate Evaporate the solvent to give 1,3,6 'triN benzyloxycarbonyl 5 epi azido 5 deoxy 2 " O 15 benzoyl 3 ",4 " N,O carbonyl 2 ' desaminosisomicin.

2 5-Epi-azido-5-deoxy-2 '-desaminosisomicin Reflux a solution of I gm 1,3,6 ' tri N benzyloxycarbonyl 5 epi azido 5 deoxy 2 " O benzoyl 3 ",4 " N,O carbonyl 2 ' desaminosisomicin in 25 ml 1:1 dioxane/water and 25 ml 10, sodium hydroxide for 24 20 hours Evaporate the solution to dryness, dissolve the residue in 10 ml water and neutralize with acetic acid Evaporate the solution, take up the residue in 5 ml.

water and pass through 20 gm of an Amberlite IRC-50 (H form) resin column, wash the column with 200 ml water and then with 100 ml IN ammonium hydroxide Collect the ammonium hydroxide eluate and evaporate to a residue 25 Freeze dry the residue, then chromatograph on a 25 gm silica gel column, eluting with chloroform:methanol:7 W ammonium hydroxide ( 2:1:1) to obtain 5 epi azido 5 deoxy 2 ' desaminosisomicin.

B 5-Epi-Amino-5-Deoxy-2 '-Desamninosisomicin Dissolve the product of step Al) in a mixture of 10 ml of tetrahydrofuran and 30 ml of liquid ammonia Slowly add 2 gm of sodium to the stirred mixture and continue to stir at -40 C for 2 hours Allow the ammonia to evaporate at room temperature overnight Dissolve the resultant residue in 25 ml of water and heat to C overnight Cool the solution and absorb on Amberlite IRC-50 (H ) resin and elute the product with 500 ml of IN ammonium hydroxide Concentrate the 35 ammonium hydroxide eluate under high vacuum to give an oily product.

Chromatograph this material on 50 gm of silica gel using chloroform:methanol:15 / ammonium hydroxide ( 2:1:1) to give 5 epi amino deoxy 2 ' desaminosisomicin.

EXAMPLE 9 40

I 6-Azidomethyl-5,6-Dihydropyran Dissolve 4 gm of 6 hydroxymethyl 5,6 dihydropyran and 10 5 gm of triethylamine in 100 ml of anhydrous dichloromethane Cool the solution and add 5.2 gm methanesulfonylchloride in 20 ml of dichloromethane, and stir the reaction for 5 hours Partition the reaction between water and chloroform, dry the 45 chloroform layer over Mg SO 4 and then evaporate to a residue in vacuo Dissolve the residue in 100 ml of dimethylformamide and add 14 gm of sodium azide and stir at 50 C for 60 hours Filter the reaction mixture, pour the filtrate into ice-water and extract with ether Evaporate the ether extracts in vacuo Take up the residue in hexane and wash with water Dry the organic layer and evaporate in vacuo, and 50 chromatograph the residue on silica gel using 3 , acetone in hexane as the eluant to obtain 6 azidomethyl 5,6 dihydropyran, pmr (CDCI 3) 8 ( 1 19 ( 4 H, m, ring CH 2), 3 35 ( 2 H, d, J= 5 5 Hz, CH 2-N 3), 4 0 (IH, m, H-5), 4 75 (IH, m, H-2), 6 4 (IH, dt, J= 1 5, 1 5, 6 5, H-I); vmax (film) 2100, 1650, 1475, 1130 cm-'.

2 6-Azido-6-Deoxy-3,4-Di-O-Benzyl-D-Glucal 55 Suspend 5 16 gm of sodium hydride in 50 ml of anhydrous dimethylformamide Maintain the temperature at O C and add 12 4 ml of benzyl chloride, then 4 65 gm of 6 azido 6 deoxy D glucal in 17 ml of dimethylformamide Stir for 4 hours and then add 15 ml of ethanol, after a further 10 minutes, pour the reaction mixture on ice and then extract with chloroform 60 1,577,540 Wash the extracts with water, then dry over Mg SO 4 and then reduce in vacuo.

Chromatograph the residue on silica gel using 25 O ethyl acetate in benzene as the eluant to obtain 6 azido 6 deoxy 3,4 di O benzyl D glucal, lal 1 + 34 3 (Et OH); pmr (CDDI 3) 83 52 ( 2 H, m, -CH 2 N 3), 4 6 ( 4 H, m, Ar-CH 2-), 6 38 IH, dd, J= 1 5, 6 Hz, H-I); vmax (CHCI 3) 2100, 1640 cm- 5 3 6 Acetamido 6 Deoxy 3,4 di O Benzyl D Glucal Stir a suspension of 760 mg of lithium aluminum hydride in 50 ml of tetrahydrofuran and to this add dropwise a solution of 298 gm of 6 azido 6 deoxy 3,4 di O benzyl D glucal in 30 ml of tetrahydrofuranHeat the mixture under reflux for 4 hours, cool and add wet ether, filter evaporate the 10 filtrate to a residue To the residue add 3 ml of acetic anhydride and 10 ml of distilled pyridine, stir for 2 hours and remove the solvent, crystallize the residue in chloroform/hexane to obtain 6 acetamido 6 deoxy 3,4 di O benzyl D glucal lal 26-41 8 (Et OH); pmr (CDCI 3) 81 86 ( 3 H, s, COCH 3), 3 65 ( 2 H,m, -CH 2-N), 4 6 ( 2 H, m, Ar CH 2), 4 75 ( 2 H, m, Ar-CH 2), 4 9 ( 1 H, dd, J= 3, 6 Hz, 15 H-2), 6 45 (IH, dd, J=l 5, 6 Hz, H-I); vmax (nujol) 3250, 1640 cm-'.

4 6-C-Methyl-3,4-Di-O-Benzyl-D-Glucal (Mixture of Diastereomeric Alcohols) Dissolve 1 gm of 6 azido 6 deoxy 3,4 di O benzyl D glucal in 250 ml of dichloromethane and photolyse for 18 hours with a 450 W mercury lamp using a Pyrex (Registered Trade 20 Mark) filter Evaporate the solution and take up the residue in 50 ml of dry ether.

To this residue add dropwise, with cooling, 10 ml of 1 3 molar equivalents of methyl magnesium bromide in ether Add water and separate out the ether layer.

Wash the ether layer with water and dry over Mg SO 4 Evaporate the ether and chromatograph the residue on silica gel using benzene as the eluant to obtain the 25 mixture of diastereomeric alcohols 6 C methyl 3,4 di O benzyl D glucal.

6 Azido 6 Deoxy C Methyl 3,4 Di O Benzyl D Glucal (Mixture of Diastereomers) Dissolve 500 mg of 6 C methyl 3,4 di O benzyl D glucal in 3 5 ml of pyridine, cool to O C and add 1 8 molar 30 equivalents of p-toluenesulfonyl chloride Let the mixture stand at 4 C for 18 hours, then add to ice-water Extract with ether, wash the ether extracts with water and dry over Mg SO 4 Evaporate off the solvent and dissolve the residue in 5 ml of dimethylformamide Add 0 5 gm of sodium azide and stir for 18 hours at 60 C.

Add the reaction mixture to water and extract with ether Dry the extracts over 35 Mg SO 4 and then reduce to a residue and chromatograph the resultant residue on silica gel using benzene at the eluant to obtain diastereomeric mixture of 6 azido 6 deoxy 6 C methyl 3,4 di O benzyl D glucal.

6 6 Amino 6 N Methyl 6 N Benzyloxycarbonyl 3,4,6 Tri deoxy D Glucal 40 Dissolve 5 gm of 6 azidomethyl 5,6 dihydropyran in 50 ml of ethanol, add 1 gm of sodium borohydride and heat at reflux for 24 hours Remove the solvent in vacuo and extract the residue with chloroform Wash the chloroform extract with water and dry over Mg SO 4 Evaporate the chloroform and dissolve the residue comprising 6 aminomethyl 5,6 dihydropyran in 100 ml of aqueous 45 methanol ( 1:1) Add 3 gm of sodium carbonate and 5 gm of benzyl chloroformate and stir the reaction for 2 hours Add 200 ml of water and extract with chloroform.

Dry the chloroform extracts and then evaporate to a residue Dissolve the residue comprising 6 benzyloxycarbonylaminomethyl 5,6 dihydropyran in '50 ml.

dimethylformamide, add 1 gm of sodium hydride ( 50 % dispersion) and 0 5 ml of 50 methyl iodide Stir the reaction mixture for 2 hours and then destroy the excess methyl iodide with acetic acid Add water and extract with chloroform, evaporate the chloroform and chromatograph the resulting residue on a silica gel column eluting with 1 % methanol in chloroform to obtain 6 amino 6 N methyl 6 Nbenzyloxycarbonyl 3,4,6 trideoxy D glucal 55 7 6 Azido 3 O Benzyl 4,6 Dideoxy D threo hex I enopy ranose Dissolve 33 2 gm of 1,2,3,6 tetra O acetyl 4 deoxy i D xvlo hexopyranose in 300 ml of chloroform (ethanol-free) Add a solution 40 gms of titanium tetrabromide in 100 ml of chloroform (ethanol-free) and reflux the 60 1.577-540 mixture for 2 hours Cool the chloroform solution and wash successively with water, aqueous sodium bicarbonate, water, and then dry the chloroform solution over Mg SO 4 Evaporate the chloroform to dryness to obtain 2,3,6 tri O acetyl 4 deoxy a D xylo hexopyranosyl bromide.

Dissolve 33 gm of the bromide in 300 ml of aqueous acetic acid ( 1:1) and add 5 portionwise 60 gin of zinc at O C, then stir for 1 5 hours Add ethyl acetate and filter the slurry and wash the solids with ethyl acetate Combine the filtrates and wash successively with water, aqueous sodium bicarbonate, water and then dry over Mg SO 4 Evaporate the ethyl acetate solution to a residue and chromatograph on a silica gel column ( 110 x 2 5 cm) using 20 / acetone in hexane as the eluant to 10 obtain 3,6 di O acetyl 4 deoxy D threo hex I enopyranose.

Dissolve 18 gm of the acetate in 400 ml of concentrated ammonium hydroxide and let stand at 25 C for 24 hours Evaporate to dryness to obtain 4 deoxy D threo hex I enopyranose.

Dissolve 10 gm of the alcohol and 15 4 gm of tosyl chloride in 100 ml of dry 15 pyrydine and let the solution stand at 25 C for 17 hours Add ethanol and evaporate the solution to a residue and then azeotrope with toluene.

Chromatograph the residue on a silica gel column ( 110 x 2 5 cm) using 10 % acetone in hexane as the eluant to obtain 4 deoxy 6 O tosyl D threo hex I enopyranose 20 Dissolve 15 gm of the tosyl compound in 300 ml of dry dimethylformamide.

Add 10 6 gm of sodium azide and stir the mixture for 24 hours at 25 C Pour the reaction mixture into ether and wash the ether layer with water, dry over Mg SO 4 and then evaporate to a residue to obtain 6 azido 4,6 dideoxy D threo hex I enopyranose 25 Dissolve 7 5 gm of the azide in 150 ml of dry dimethylformamide and add 2 5 gin of sodium hydride Stir the mixture at 25 C for 1 hour, then add 10 gm of benzylbromide and stir the mixture at 25 C for 6 hours Add a 20 % solution of methanol in ether and filter off the solids and wash with chloroform Combine the filtrates and pour into water and extract with chloroform and dry over Mg SO 4 30 Evaporate the chloroform extract and chromnatograph the resultant residue on a silica gel column ( 110 x 2 5 cm) using 5 %,, acetone in hexane as the eluant to obtain 6 azido 3 O benzyl 4,6 dideoxy D threo hex I enopyranose.

8 1,3,3 ' tri N Benzyloxycarbonyl 1 N Ethylgaramine Dissolve 10 47 gin of 1-N-ethylgaramine in 300 ml of 60 % aqueous methanol 35 Add 15 9 gm of sodium carbonate and to the reaction add (dropwise) 25 6 gm of benzyloxycarbonyl chloride Stir the reaction mixture for 2 5 hours and then add 9.6 gm of sodium carbonate and 15 3 gm of benzyloxycarbonyl chloride Stir the reaction for 3 hours and then pour the solution into water Extract with chloroform, wash the chloroform extracts with water and dry over Mg SO 4 Concentrate the 40 chloroform solution to a small volume and add it with stirring to etherhexane ( 1:1).

Filter the resultant solid to obtain 1,3,3 ' tri N benzyloxycarbonyl 1 N ethylgaramnine, lal 6 + 76 2 (Me OH); pmr (CDCI 3) 81 05 ( 6 H, m, -CH 2-CH 3 and C-CH 3), 3 02 ( 3 H, s, N-CH 3), 5 05 ( 6 H, m, Ar-CH 2), 7 3 ( 15 H, m, aromatic).

9 5,2 ',4 'tri O Acetyl 1,3,3 ' tri N Benzyloxycarbonyl I N 45 Ethylgaramine Dissolve 18 0 gm of 1,3,3 ' tri N benzyloxycarbonyl 1 N ethylgaramnine in 300 ml of pyridine Stir the solution at O C and add, dropwise, 5 8 gm of 2,2,2 trichloroethyl chloroformate, then let the reaction stand at room temperature for 18 hours Then add the solution to ice-water and extract the 50mixture with ethyl acetate Wash the ethyl acetate extracts successively with water, dilute HCI, water and then dry the ethyl acetate over Mg SO 4 Concentrate the ethyl acetate solution and add it, with stirring, to hexane-ether ( 1:1) Collect the 4 O trichloroethoxycarbonyl derivative by filtration and dissolve in a mixture of 160 ml of glacial acetic acid, 83 75 ml of acetic anhydride and 9 25 ml of 55 concentrated HCI and heat on a steam bath for 3 5 hours Cool the solution and pour into ice-water Extract with ethyl acetate and wash the extracts with water, dilute sodium bicarbonate solution, water and then dry over Mg SO 4 Evaporate the solvent and dissolve the residue comprising 5,2 ',4 ' tri O acetyl 1,3,3 ' tri N benzyloxycarbonyl I N ethyl 4 trichloroethoxycarbonylgaramine in 60 700 ml of 90 , acetic acid in water Add 100 gm of zinc dust and stir for 5 hours, then add an additional 30 gm of zinc and stir a further 16 hours Filter off the unreacted zinc and reduce the filtrate in vacuo Take up the residue in ethyl acetate, 1,577,540 wash with water and dry over Mg SO 4 Chromatograph on a silica gel column using l Y, methanol in chloroform as the eluant to obtain 5,2 ',4 ' tri O acetyl 1,3,3 ' tri N benzyloxycarbonyl I N ethylgaramine, la I 6 + 53 5 (Me OH) ; pmr (CDCI 3) 81 12 ( 3 H, t, J= 6 5 Hz, -CH 2 CH 3), 1 34, 1 4 ( 3 H, CCH 3), 2 88 ( 3 H, s, N-CH 3), 5 12 ( 6 H, m, Ar-CH 2), 7 35 ( 15 H, m, aromatic) 5 The present invention includes within its scope pharmaceutical compositions comprising the novel 2 '-unsubstituted derivatives of 4,6 di O (aminoglycosyl) 1,3 diaminocyclitols with a compatible, pharmaceutically acceptable carrier, or coating Also included within the invention is the method of eliciting an antibacterial response in a warm-blooded non-human animal having a 10 susceptible bacterial infection which comprises administering to said animal a nontoxic, antibacterially effective amount of a member selected from the group consisting of a 2 '-unsubstituted derivative of a 4,6 di O (aminoglycosyl) 1,3 diaminocyclitol having antibacterial activity.

The 2 '-unsubstituted derivative of 4,6 di O (aminoglycosyl) 1,3 15 diaminocyclitols of this invention and the non-toxic pharmaceutically acceptable acid addition salts thereof are broad spectrum antibacterial agents which, advantageously, exhibit activity against organisms, particularly gramnegative organisms Furthermore, it has been found that removal of the 2 '-amino group confers activity against the 2 '-N-acetylating strains and in addition, these 20 compounds, as well as those in which the 2 '-hydroxyl has been removed, have increased potency against the 3-N-acetylating strains Thus, the compounds of this invention can be used alone, or in combination with other antibiotic agents to prevent the growth, or reduce the number of bacteria in various environments The activity of the 2 '-unsubstituted4,6 di O (aminoglycosyl) 1,3 25 diaminocyclitols against gram-negative bacteria renders them useful for combating infections caused by gram-negative organisms, e g species of E coli, Klebsiella, Proteus, Providencia, Salmonella, Serratia and Pseudomonas; and, furthermore, for combating infections caused by gram-positive organisms, e g species of Staphylococcus, Streptococcus and BSubtilis 30 The compounds, e g 2 '-deoxygentamicin B and 2 '-desaminosisomicin have veterinary applications, particularly in the treatment of mastitis in cattle and Salmonella-induced diarrhea in domestic animals such as the dog and the cat The compounds may also be used, for example, to disinfect laboratory glassware, dental and medical equipment contaminated with the gram-positive organism, 35 Staphylococcus aureus, or other bacteria.

In general, the dosage administered of the 2 '-unsubstituted 4,6 di O (aminoglycosyl) 1,3 diaminocyclitols will be dependent upon the age and weight of the animal species being treated, the mode of administration, and the type and severity of bacterial infection being prevented, or reduced 40 The 2 ' unsubstituted 4,6 di O (aminoglycosyl) 1,3 diaminocyclitols and the pharmaceutically acceptable acid addition salts thereof may be administered orally They may also be applied topically in the form of ointments, both hydrophilic and hydrophobic, in the form of lotions which may be aqueous, nonaqueous, or other emulsion type, or in the form of creams or gells 45 Pharmaceutical carriers useful in the preparation of such formulations will include, for example, such substances as water, oils, fats, waxes, polyesters, alcohols and polyols.

In general, the topical preparations preferably contain from 0 1 to 3 0 gms of the compounds of this invention per 100 gms of ointment, creams or lotion The topical preparations are usually applied gently to lesions from 2 to 5 times a day.

For oral administration the antibacterials of this invention may be compounded in the form of tablets, capsules or elixirs, or may even be admixed with animal feed It is in these dosage forms that the antibacterials are most effective for treating bacterial infections of the gastrointestinal tract which 55 infections cause diarrhea They are also useful for pre and post-operative gut sterilization.

The antibacterials of this invention may be utilized in liquid form such as solutions and suspensions for otic and opthalmic use and may also be administered parenterally via intramuscular, intravenous, subcutaneous and intrasternal 60 injection The injectable solution, or suspension will usually be administered at from 1 mg to 15 mgs of antibacterial per kilogram of body weight per day divided into 2 to 4 doses.

1.577540 The following formulations are to exemplify some of the dosage forms in which the antibacterial agents of this invention may be employed:

Formulation 1 Tablet 10 mg Tab 25 mrg Tab 100 mg Tab.

2 '-Deoxygentamicin B 10 50 mg 26 25 mg 105 00 mg 5 Lactose, impalpable powder 197 50 mg 171 25 mg 126 00 mg.

Corn starch 25 00 mg 25 00 mg 35 00 mg.

Polyvinylpyrrolidone 7 50 mg 7 50 mg 7 50 mg.

Magnesium Stearate 2 50 mg 2 50 mg 3 50 mg 10 243 00 mg 232 50 mg 277 00 mg.

50, excess.

Procedure Prepare a slurry consisting of the 2 '-deoxygentamicin B, lactose and polyvinylpyrrolidone Spray dry the slurry Add corn starch and magnesium 15 stearate Mix and compress into tablets on a suitable press to the specified weight.

Formulation 2 Ointment 2 '-Deoxygentamicin B 1 0 gm.

Methyl paraben USP 0 5 gm 20 Propyl paraben USP 0 1 gm.

Petrolatum to 1000 gm.

Procedure (I) Melt the petrolatum.

( 2) Mix the 2 '-deoxygentamicin B, methyl paraben and propyl paraben with 25 about 10 , of molten petrolatum and make a slurry Mill the slurry and add to the balance of the petrolatum Cool to room temperature with agitation.

Claims (1)

1. WHAT WE CLAIM IS:-

   1 A 2 ' unsubstituted 4,6 di O (aminoglycosyl) 1,3 diaminocyclitol, wherein the 4 O aminoglycosyl is defined by one of the 30 following Formula I and II:

   R 1-C-NHT R _C-NHT 0 5 41 R 41 RI 31 I' 2 '10-31 21 I 11 wherein R and R' are each hydrogen or hydroxy; R" is hydrogen or methyl; T is hydrogen, alkyl, hydroxyalkyl or aminoalkyl said alkyl having up to 4 carbon atoms; and when the 1,3-diaminocyclitol has a 5-hydroxyl group in the equatorial 35 position, the 5-epi, 5-deoxy, 5-epi-azido-5-deoxy, 5-epi-amino-5-deoxy derivatives thereof; the l-N-X-derivatives of the foregoing, wherein X is a substituent -CH 2-Y or -C-Y wherein Y is hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, hydroxyalkyl, 40 aminoalkyl, mono or dialkylaminoalkyl, aminohydroxyalkyl, mono or dialkylaminohydroxyalkyl, phenyl, benzyl or tolyl, said aliphatic radicals having up to 8 carbon atoms and when substituted by both amino and hydroxy groups, said 1,577,540 groups are on different carbon atoms; and the pharmaceutically acceptable acid addition salts thereof.

   2 A compound of claim I wherein the 6 O (aminoglycosyl) is defined by the following Formula III R 2 0 NH 15 4 X 0OH wherein R, is hydrogen or methyl, R 2 is hydrogen or hydroxymethyl, R 3 is hydrogen or hydroxy and R 4 is hydrogen, methyl or hydroxy with the proviso that both R 3 and R 4 are not hydroxy.

   3 A compound of claim 1, Formula I wherein R and R' are hydroxy, R" is hydrogen or methyl, and T is hydrogen, selected from: 10 2 '-deoxygentamicin B, 2 '-deoxygentamicin B 1, 2 '-deoxygentamicin A 3, 2 '-deoxykanamycin A, and 2 '-desamino-Antibiotic JI-20 B 15 4 A compound of claim I, Formula I wherein R and T are hydrogen, R' is hydroxy, and R" is hydrogen or methyl, selected from:

   2 ',3 '-dideoxygentamicin B, 2 ',3 '-dideoxygentamicin B 1, 2 ',3 '-dideoxygentamicin A 3, 20 2 ',3 '-dideoxykanamycin A, and 2 '-desamino-3 '-deoxy-Antibiotic JI-20 B. A compound of claim 1, Formula I wherein R and R' are hydrogen, and R" and T are hydrogen or methyl, selected from:

   2 '-desaminogentamicin C, 25 2 '-desaminogentamicin C 1 a, 2 '-desaminogentamicin C 2, 2 '-desaminogentamicin C 2 a, 2 '-desaminogentamicin C 2 b, 2 ',3 ',4 '-trideoxykanamycin A, and 30 2 ',3 ',4 '-trideoxygentamicin A 3.

   6 A compound of claim 1, Formula II wherein R" and T are hydrogen or methyl, selected from:

   2 '-desaminosisomicin, 2 '-desaminoverdamicin, 5 2 '-desamino-Antibiotic 66-40 B, 2 '-desamino-Antibiotic 66-40 D, 2 '-desamino-Antibiotic G-52, 2 '-desamino-Antibiotic Mu-l, and 2 '-desamino-Antibiotic Mu-4 40 7 A compound of claims I and 2, wherein the 6 O (aminoglycosyl) is garosaminyl and the 1,3-diaminocyclitol is 2-deoxystreptamine.

   8 A 5-epi or 5-deoxy derivative of a compound of claim 7.

   9 A l-N-X derivative of a compound of any one of claims 3 to 8 wherein X is -CH 2-Y, Y being alkyl or aminoalkyl having up to 3 carbon atoms 45 A l-N-X derivative of a compound of any one of claims 3 to 8 wherein X is a C-Y 1.577540 substituent selected from p-amino a hydroxypropionyl, y amino a hydroxybutyryl, and S amino a hydroxyvaleryl.

   11 I N ( 3 amino a hydroxypropionyl) 2 ',3 'dideoxygentamicin B or a pharmaceutically acceptable acid addition salt thereof.

   12 1 N ethyl 2 ',3 'dideoxygentamicin B or a pharmaceutically 5 acceptable acid addition salt thereof.

   13 1 N (/p amino a hydroxypropionyl) 2 ' desaminosisomicin or a pharmaceutically acceptable acid addition salts thereof.

   14 1 N ethyl 2 ' desaminosisomicin or a pharmaceutically acceptable acid addition salt thereof 10 A process for preparing a compound as claimed in claim 1, wherein a 6 O (aminoglycosyl) 1,3 diaminocyclitol having an unprotected 4-hydroxyl group and being suitably protected at other positions is condensed with a suitably protected glycal in the presence of an acid catalyst; any protecting groups are removed by known methods (as defined herein) and wherein, if desired, a 5epi, 5 15 deoxy, 5 epi azido 5 deoxy or 5 epi amino 5 deoxy and/or a I-N-X and/or a 6 '-N-T' derivative is prepared by known methods with X being as defined in claim I and T' being the same as T in claim 1 except for hydrogen; the last step being followed by isolating the product as such or as a pharmaceutically acceptable acid addition salt 20 16 Process of claim 15, wherein the acid catalyst is selected from sulphonic acids, sulphonic acid resins, sulfuric acid, phosphoric acid, trifluoroacetic acid and sulfamic acid.

   17 Process of claim 16, wherein the acid catalyst is p-toluene sulphonic acid.

   18 Process of any one of claims 15 to 17, wherein as the 6 O 25 (aminoglycosyl) 1,3 diaminocyclitol garamine having an unprotected 4hydroxyl group and having allamino groups and all other hydroxyl groups protected is used either as such or as the 1-N-X derivative thereof with X being -CH 2 Y and Y is as defined in claim 1.

   19 A pharmaceutical composition comrising as active ingredient at least one 30 compound as claimed in claim 1, together with a pharmaceutical carrier or excipient.

   A composition of claim 19 in the form of dosage units.

   21 Process for preparing a pharmaceutical composition of claims 19 and 20 wherein the active ingredient is admixed with a pharmaceutical carrier or 35 excipient.

   22 A method of eliciting an antibacterial response in a warm-blooded nonhuman animal having a susceptible bacterial infection, which comprises administering to said animal a non-toxic, antibacterially effective amount of a compound as claimed in claim 1 40 23 A process of claims 15 to 18 substantially as herein described in any of the Examples I to 8.

   24 A compound of claims I to 14 whenever prepared by the process of claims to 18.

   25 A pharmaceutical composition of claim 19 or 20 substantially as herein 4 described with specific reference to the Formulations.

   26 A pharmaceutical composition whenever obtained by the process of claim 21.

   MATHYS & SQUIRE, Chartered Patent Agents, 10, Fleet Street, London EC 4.

   Agents for the Applicants.

   Printed for Her Majesty's Stationery Office, by the Courier Press, Leamington Spa 1980 Published by The Patent Office, 25 Southampton Buildings, London WC 2 A l AY, from which copies may be obtained.

   1,577,540